Nanobiotechnology for the Treatment of Mantle Cell Lymphoma

纳米生物技术治疗套细胞淋巴瘤

• 批准号: 7746996
• 负责人: TRUDY M FORTE
• 金额: $ 10.94万
• 依托单位: LYPRO BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746996
• 关键词: Acute Promyelocytic Leukemia; Aggressive course; Amino Acid Sequence; Animal Model; Antibodies; Antigens; Apolipoprotein A-I; Apolipoproteins; Apoptosis; Autophagocytosis; B lymphoid malignancy; B-Lymphocytes; Binding; Biological; C-terminal; CD20 Antigens; Caspase; Categories; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Chemosensitization; Chimera organism; Chimeric Proteins; Code; Controlled Study; Cultured Cells; Disease; Disease remission; Dose; Drug Delivery Systems; Effectiveness; Escherichia coli; Evaluation; Exhibits; Exposure to; Flow Cytometry; Gene Targeting; Human; Induction of Apoptosis; Investigation; Juvenile Myelomonocytic Leukemia; Kaposi Sarcoma; Lipids; Lymphoma; MS4A1 gene; Malignant Neoplasms; Mantle Cell Lymphoma; Measurement; Mediating; Methods; Modeling; Neuroblastoma; Non-Hodgkin' s Lymphoma; Nuclear Hormone Receptors; Oncogenes; Parents; Plasmid Cloning Vector; Production; Property; Protein Engineering; Proteins; RXR; Reaction Time; Reactive Oxygen Species; Recombinant Fusion Proteins; Research; Research Design; Research Personnel; Resistance; Retinoic Acid Receptor; Retinoids; Role; Surface; Time; Tissues; Treatment Protocols; Tretinoin; Western Blotting; base; cancer therapy; cell growth; cell type; high risk; in vivo; member; nanobiotechnology; nanoscale; novel; novel strategies; outcome forecast; particle; public health relevance; targeted delivery

DESCRIPTION (provided by applicant): Mantle cell lymphoma (MCL) is a B-cell malignancy that is characterized by dysregulation of various oncogenes. Building on evidence that retinoic acid and its derivative, all trans retinoic acid (ATRA), are useful agents that potentiate apoptosis or anti-proliferative effects, it is proposed to pursue a strategy of targeted delivery of ATRA to MCL cells in culture. Using a novel delivery vehicle wherein ATRA is solubilized in nanoscale, protein stabilized lipid particles, termed nanodisks (ND), protein engineering methods will be employed to target ATRA-ND to the CD20 antigen present on the surface of B lymphocytes. This will be achieved by construction of a single chain variable antibody (scFv) apolipoprotein (apo) fusion protein. It is hypothesized that 1-CD20 scFv7apoA-I fusion protein will be capable of forming ND while retaining the antigen recognition properties of the parent scFv. Recombinant fusion protein will be expressed in E. coli, isolated and characterized. The ability of the chimera to associate with lipid and induce formation of ND will be determined while the antigen recognition properties of the 1-CD20 scFv portion of the fusion protein will be evaluated by Western blot and flow cytometry. ND prepared with wild type apoA-I will serve as control for these studies. In a second aim the effect of 1-CD20 scFv7apoA-I ATRA-ND on MCL cells in culture will be assessed. It is hypothesized that retinoid containing, targeted ND, will display enhanced induction of apoptosis in cell culture models of MCL. Different MCL cell lines will be employed in studies designed to evaluate targeting efficiency, concentration effectiveness and cell viability following exposure to CD20 targeted scFv7apoA-I ATRA-ND. Cultured cells will be exposed to control ATRA-ND and 1-CD20 scFv7apoA-I ATRA-ND followed by measurements of cell viability, apoptosis and autophagy. Dose- response and time course studies will be conducted to define optimal conditions. Studies will also be performed with ND harboring related synthetic and natural retinoids. The results of these studies will expand the potential of ND mediated drug delivery by demonstrating cell / tissue specific targeting, providing a framework for in vivo studies of targeted ND in animal models of MCL. PUBLIC HEALTH RELEVANCE: Despite progress made in the broad category of lymphomas, mantle cell lymphoma (MCL) remains a poorly treated disease with median survival time of approximately 3 to 4 years. New therapy regimens have increased the complete remission rate but they have done little to change overall survival. Research proposed herein will evaluate the effectiveness of targeted drug payload delivery to cultured MCL cells. These studies will establish a novel approach with broad applicability, establishing targeted nanodisks as a platform that can be used with other forms of cancer, potentially decreasing the public burden associated with this disease.

描述（由申请人提供）：地幔细胞淋巴瘤（MCL）是B细胞恶性肿瘤，其特征是各种发育不良的失调。在证据表明视黄酸及其衍生物（所有反式视网膜酸）（ATRA）的基础上是有用的药物，可以增强凋亡或抗增殖作用，因此建议采取针对培养中MCL细胞靶向递送ATRA的策略。使用一种新型的递送载体，其中ATRA在纳米级中溶解，蛋白质稳定的脂质颗粒（称为纳米虫（ND））将采用蛋白质工程方法将ATRA-ND靶向于B淋巴细胞表面上存在的CD20抗原。这将通过构建单链可变抗体（SCFV）载脂蛋白（APO）融合蛋白来实现。假设1-CD20 SCFV7APOA-I融合蛋白将能够在保留父scFV的抗原识别特性的同时形成ND。重组融合蛋白将在大肠杆菌中表达，分离和表征。将确定融合蛋白1-CD20 SCFV部分的抗原识别特性，将通过蛋白质印迹和流动细胞仪评估嵌合体与脂质和诱导ND形成的能力。使用野生型Apoa-I制备的ND将作为这些研究的控制。在第二个目标中，将评估1-CD20 SCFV7APOA-I ATRA-ND对MCL细胞在培养中的影响。假设含有靶向ND的类视黄素将显示出MCL细胞培养模型中凋亡的诱导增强。旨在评估靶向效率，浓度有效性和细胞活力后，将采用不同的MCL细胞系。培养的细胞将暴露于控制ATRA-ND和1-CD20 SCFV7APOA-I ATRA-ND，然后测量细胞活力，凋亡和自噬。将进行剂量反应和时间课程研究以定义最佳条件。研究还将使用ND携带相关的合成和天然类视角素进行。这些研究的结果将通过证明细胞 /组织特异性靶向来扩大ND介导的药物递送的潜力，从而为MCL动物模型中靶向ND的体内研究提供了一个框架。 公共卫生相关性：尽管在广泛的淋巴瘤类别中取得了进展，但地幔细胞淋巴瘤（MCL）仍然是一种治疗良好的疾病，中位生存时间约为3至4年。新的疗法方案提高了完整的缓解率，但它们几乎没有改变整体生存率。本文提出的研究将评估靶向药物有效载荷到培养的MCL细胞的有效性。这些研究将建立一种具有广泛适用性的新型方法，建立靶向纳米风险作为一个可以与其他形式的癌症一起使用的平台，并可能减轻与这种疾病相关的公共负担。